Chilli peppers--a review on tissue culture and transgenesis.

Chili pepper is an important horticultural crop that can surely benefit from plant biotechnology. However, although it is a Solanaceous member, developments in plant cell, tissue, and organ culture, as well as on plant genetic transformation, have lagged far behind those achieved for other members of the same family, such as tobacco (Nicotiana tabacum), tomato (Lycopersicon esculentum), and potato (Solanum tuberosum), species frequently used as model systems because of their facility to regenerate organs and eventually whole plants in vitro, and also for their ability to be genetically engineered by the currently available transformation methods. Capsicum members have been shown to be recalcitrant to differentiation and plant regeneration under in vitro conditions, which in turn makes it very difficult or inefficient to apply recombinant DNA technologies via genetic transformation aimed at genetic improvement against pests and diseases. Some approaches, however, have made possible the regeneration of chili pepper plants from in vitro-cultured cells, tissues, and organs through organogenesis or embryogenesis. Anther culture has been successfully applied to obtain haploid and doubledhaploid plants. Organogenic systems have been used for in vitro micropropagation as well as for genetic transformation. Application of both tissue culture and genetic transformation techniques have led to the development of chili pepper plants more resistant to at least one type of virus. Cell and tissue cultures have been applied successfully to the selection of variant cells exhibiting increased resistance to abiotic stresses, but no plants exhibiting the selected traits have been regenerated. Production of capsaicinoids, the hot principle of chili pepper fruits, by cells and callus tissues has been another area of intense research. The advances, limitations, and applications of chili pepper biotechnology are discussed.

In vitro chili pepper biotechnology

Plant Tissue Culture Procedure - Background

Two S-(2-aminoethyl)L-cysteine (AEC) resistant lines were isolated by screening mutagenized protoplasts from diploid N. sylvestris plants. Both lines accumulated free lysine at levels 10 to 20-fold higher than in controls. Lysine overproduction and AEC-resistance were also expressed in plants regenerated from the variant cultures. A feedback insensitive form of dihydrodipicolinate synthase (DHPS), the pathway specific control enzyme for lysine synthesis, was detected in callus cultures and leaf extracts from the resistant lines. Aspartate kinase (AK), the other key enzyme in the regulation of lysine biosynthesis, was unaltered in the mutants. Crosses with wild type plants indicated that the mutation conferring insensitivity to feedback in DHPS, with as result overproduction of lysine and resistance to AEC, was inherited as a single dominant nuclear gene.

Lysine overproducer mutants with an altered dihydrodipicolinate synthase from protoplast culture of Nicotiana sylvestris (Spegazzini and Comes).

Protoplast-derived colonies of haploid N. plumbaginifolia were selected for by chlorate resistance in media supplemented with casamino acids. Eighty resistant lines were confirmed by a second passage on a higher concentration of chlorate. Frequency of spontaneous mutation ranged from 10−5 to 10−6. Fifty of the resistant lines could be regenerated into plants, and 30 were characterized biochemically. Ninety percent were fully deficient for nitrate reductase activity. The lines were further tested for xanthine dehydrogenase activity and subsequently classified as defective in the apoenzyme (nia type, 26 lines) or the cofactor (cnx type, 4 lines). Two groups had been identified up until now within the cnx type by growth tests on high concentrations of molybdate supplied to the medium. Nitrate reductase deficiency was stably and continously expressed in both variant cell cultures and regenerants. Genetic analysis demonstrated that nitrate reductase deficiency was inherited as a single recessive nuclear gene.

Regeneration of fully nitrate reductase - deficient mutants from protoplast culture of Nicotiana plumbaginifolia (Viviani).

High and Reproducible Plating Efficiencies of Protoplasts isolated from in vitro Grown Haploid Nicotiana sylvestris SPEGAZ. et COMES

Changes in rectal and mean skin temperature in response to suggested heat during hypnosis in man.

Parameters computed from electrocardiographic recordings (mean frontal QRS axis, â QRS, positive (R-S) difference in lead V1, incidence of atypical conduction pattern in V1) were compared: (1) in two populations residing at the same altitude (3 800 m) but in different geographical sites: Aymaras in Bolivia and Tibetans in Nepal, (2) in three groups of Bolivians dwellers, ethnically similar and fully acclimatized, at three altitudes (4 780 m, 3 800 m, 400 m). This work involved 661 subjects. Results: (a) The mean â QRS value in highlanders is shifted to the right when compared to that of lowlanders: the right axis deviation increases with altitude, (b) The mean â QRS value is identical in Bolivian and Tibetan groups living at the same altitude, (c) The axis deviates to the left with aging in all the environmental conditions. This migration is accompanied by a lower incidence of positive (R-S) difference in adults compared to younger subjects, (d) The mean â QRS value of the females is always situated to the left of that males for all age groups. This difference receives a possible confirmation by the lower incidence of atypical complexes in V1 in females, (e) The present values of â QRS as well as others found in the litterature and those of mean pulmonary arterial pressure reported by different authors have been plotted, both as a function of elevation: The two relationships can be described by two linear functions with a point of intersection. Such points suggest an altitude threshold above which a further decrease in barometric pressure results in marked cardiovascular responses. They are both located in the vicinity of 2 500 m.

Jacques Durand

Papers

High and Reproducible Plating Efficiencies of Protoplasts isolated from in vitro Grown Haploid Nicotiana sylvestris SPEGAZ. et COMES

Changes in rectal and mean skin temperature in response to suggested heat during hypnosis in man.

Electrocardiographic observations in high altitude residents of Nepal and Bolivia.

Heat exchange and energetic balance during exercise in highlanders and lowlanders at sea level and high altitude